The objective of this proposal is to discover components of the ambient temperature perception pathway in Brachypodium distachyon. This will help us understand the way in which crop plants perceive and respond to changes in ambient temperature, including climate change. A gene expression atlas for the major ambient temperature responses in Brachypodium will be created using microarrays, to identify genes that show a continually variable expression across the ambient (i.e. non-stress inducing) temperature range. These experiments will be compared to data from Arabidopsis to identify the level of conservation in temperature perception pathways between monocots and dicots. The promoters of temperature responsive genes identified through this analysis will be fused to the firefly Luciferase gene to develop ambient temperature reporter lines. Selected lines will be used in forward genetic screens (using fast neutron irradiation mutagenesis) to identify key components of the Brachypodium temperature sensing pathway. This analysis will be complemented by functional characterisation of temperature sensing genes identified in parallel screens in Arabidopsis that have been carried in the Wigge lab as well as using the expression atlas data. These reverse genetics approaches will use RNA interference technology to knock-down the activity of the genes of interest. The phenotypes of these plants will be assessed to determine the direct role of each candidate within the temperature perception pathway. Taken together, this project aims to provide novel information about the way in which our major crop plants will respond to changes in ambient temperature by continuing the development of Brachypodium as a model organism for temperate cereals, such as wheat and barley, and identifying genetic components within the monocot temperature perception pathway.
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